Use of {60 ,{62 -poly-(aspartic acid)-hydroxyalkylamides as a plasma expander

ABSTRACT

A method for the treatment of a patient with a diminished plasma volume level caused by microcirculatory impairment due to septic or traumatic shock comprising administrating a therapeutically effective plasma expanding dose of a hydroxyalkylamide of Alpha , Beta -polyaspartic acid having the formula   wherein: R1 is hydrogen, an alkyl group, or a hydroxyalkyl group; R2 is a hydroxyalkyl group; n is the number of beta-linked residues; m is the number of alpha-linked residues, and the sum of n + m is within the range of 100 to 300 (calculated from viscosity measurements in DMF according to Doty&#39;&#39;s curve, J. Am. Chem. Soc. 78, 947, 1956) or from 75 to 550 (calculated from ultracentrifuge measurements).

United States Patent [191 Neri et al.

[ Dec. 16, 1975 1 USE OF a,B-POLY-(ASPARTIC ACID)-HYDROXYALKYLAMIDES ASA PLASMA EXPANDER [75] Inventors: Paolo Neri; Guido Antoni; FrancoBenvenuti, all of Siena, Italy [73] Assignee: Instituto Sieroterapico eVaccinogeno Toscano Sclavo S.p.A., Siena, Italy [22] Filed: Aug. 21,1974 [21] Appl. No; 499,580

Related US. Application Data [63] Continuation-in-part of Ser. No.51,771, July 2,

[30] Foreign Application Priority Data OTHER PUBLICATIONS J. ofMedicinal Chemistry, 10, 1967, Kovacs et al., pp. 904-908.

J. of Organic Chemistry, 26, 1961, Kovacs et al., pp. 1084-1091.

Advances in Protein Chemistry, Vol. XIII, 1958, Anfinsen et al., pp.419-426.

Primary ExaminerStanley J. Friedman Attorney, Agent, or FirmHaseltine,Lake & Waters [5 7] ABSTRACT A method for the treatment of a patientwith a diminished plasma volume level caused by microcirculatoryimpairment due to septic or traumatic shock comprising administrating atherapeutically effective plasma expanding dose 'of a hydroxyalkylamideof a, B-polyas partic acid having the formula wherein: R is hydrogen, analkyl group, or a hydroxyalkyl group; R is a hydroxyalkyl group; n isthe number of beta-linked residues; m is the number of alphalinkedresidues, and the sum of n m is within the range of 100 to 300(calculated from viscosity measurements in DMF according to Dotys curve,J. Am. Chem. Soc. 78, 947, 1956) or from 75 to 550 (calculated fromultracentrifuge measurements).

5 Claims, 1 Drawing Figure US. Patent Dec. 16, 1975 3,927,204

Amiira/ used /)2 ewe/M2605: male-09f These polymers (formula (I) areeasily dissolved in USE OF mB-POLY-(ASPARTIC water and are withoutelectrical charge; the choice of ACID)-HYDROXYALKYLAMIDES AS A PLASMAsuitable substitutes permits one to obtain products EXPANDER without, orwith low toxicity.

Toxicological experiments carried out on various CROSS RELATEDAPPLICATION species of animals (mouse, rat, rabbit, cat) have show Thisapplication is a continuation-in-part of copendthat products with n mfrom 100 to 300 (from Doty's ing application Ser. No. 51,771, filed July2, 1970. curve); 75 to 550 (from ultracentrifuge measurements) arewithout acute or chronic toxicity. Also they are not FIELD OF THEINVENTION 1O antigenio This invention relates to polymers, and inparticular From a pharmacological point of view, polymers of n tohydroxyalkylamides suitable as plasma expanders +m varying from 200 to250 (from Dotys curve); 250 and useful in protecting mammals from shock.to 400 (from ultracentrifuge measurements) have shown greater efficacyin tests for protection against BRIE,- DESCRIPTION OF THE DRAWINGhaemorrhagic shock in animals, though preparations of The sole FIGURE ofthe drawing is a graphical repren m from 150 to 300 (from Dotys curve);150 to 550 sentation of the relation of bleeding volume index with (fromultracentrifuge measurements) have shown some respect to the degree ofpolymerization of the comefficacy. Both plasma volume and bleedingvolume pound of the invention. index (B.V.I.) have been determined.

' The results of this last test carried out with a,,8-poly- SUMMARY OFTHE INVENTION DL-aspartic hydroxyethylamide, for different degreesa,,B-Poly-(aspartic acid) hydroxyalkylamides of the of polymerizationare shown in the graph in the atpresent invention are embraced by thefollowing fortached drawing. In this graph the value of n m is mula (I):shown on the abscissa and the value of B.V.I. on the CONR,R2

H2NCHCO NH-CH'CH2-CO CONR,R2 cl-l H2 CONR R CH NHCHCOOH comma NH(l'H-CO.

n v m ordinate. The test for the determination of the bleeding Wheremi 115 yd g Or an alkyl g p a Y volume index was carried out in the male catby bleedy y g hpt 2 15 y y y g p; n 15 the hllming 2 ml/kg/ 3 minutesand with a rate of intravenous ber of B- feSldueS, and m 1$-thehumber of(I.V.) infusion of 3% solution in saline of 2 ml/kg/milinked residues.The sum of n r n (polymerization 40 mute, degree of polymer) wascalculated either from viscosity Th 3% l i f g l gb i hydroxyemeasurements in N,N-dimethylformamide according to h l id h b comparedwith the 6% Dextran Dotyvs curve Chem See 78, 947, 1956) of solutionthat is normally commercially available. From from hltt'aeehh'ithgetheasutemehte n m lies within this comparison, it results-that thesolution of a,B-polythe range of 100 to 300 (from y 75 to 550DL-aspartic hydroxyethylamide does not alter the pa- (ffOm l rif gmeasurements) Compounds of rameter of haemagglutination, as happens withDextran formula I herei the VOllle 0f heS Within the solutions; theincrease in erythrocyte sedimentation range of 200 to 250 (from y 250 to400 rate (E.S.R.) caused by the new product is notably less (fromultracentrifilge measurements) e of Particular than that caused byDextran; Dextran solution shows vahle- Compounds of formula I includethe D, L and some antigenicity. With regard to efficacy, the two DLisomeric forms. The compounds of formula I are products proved l,

Synthesized y the reaction of y p ye pe Therefore, thehydroxyalkylamides of a,/3-polyacid formula of shltable degree of p y haspartic acid can be usefully used as plasma expanders, Withapproximately ah equal amount of the PP P l at concentrations of 34% insaline or salt-added solue aminOEllCOhOl; the degree of polymerizationof the tions. For a more detailed qualitative and quantitative y p y pacid must be greater than that of composition, see Examples 7 and 8hereafter.

the desired polyhydroxyalkylamide, because partial DESCRIPTION OFPREFERRED EMBODIMENTS degradation takes place during the reaction.

Preferred embodiments of the invention will hereafter bevdiscussed ingreater detail with reference to the C0 C0 C0 following specificexamples. 0 a EXAMPLE 1 CH, I l a Preipa aiiloin of the formula Iwherein:

co co co V R; -l=igcn on i 30 g of anhydffipolyaspartic acid weredissolved in X g ml of dimetliylfdrmamide, and 30 ml of ethanolaminewere addea t@ the solution while stirring and keeping 3 the reactionmixture at a temperature of 2530C. After about 1 hr, the polymerobtained was precipitated by the addition of about 500 ml of chloroform.

The upper liquid solution was eliminated as much as possible bydecantation and the residual semi-solid mass was again treated withchloroform to further purify it from the ethanolamine.

The product was then dissolved in diluted HCl, and the solutionneutralized with dilute NaOH and dialyzed against running water for 72hrs and against distilled water for 48 hrs. It was then concentrated andlyophylized.

Yield: about 38 g.; Analysis: for (CA-1 N calculated: C 45.57; H 6.37; N17.71; 0 30.35; found: C 45.45; H 6.30; N 17.72; 0 30.40. It wascalculated that the degree of polymerization of this polymer is about200(Dotys curve); 250 (ultracentrifuge). The oncotic efficacy per gramof polymer in comparison with Bovine Serum Albumin is about 2.3.

EXAMPLE 2 Preparation of the product of formula 1 wherein:

l-l CH. ,OH n m 220 (Dory's curve 300 (ultracentrifuge) (averagevalues). 30 g of anhydropolyaspartic acid were dissolved in 150 ml ofdimethylformamide, and 30 ml of ethanolamine were added to the solutionwhile stirring, and the reaction mixture kept at a temperature of 2530C.After about 1 hr, g of acetic acid were added, during continual stirringfor 10 min. at room temperature, and the solution was concentrated undervacuum. The residue was taken up with water, dialyzed for 72 hrs againstrunning water and for 48 hrs against distilled water, and lyophylized.

Yield: about 42 g.; Analysis: for (C l-1 N 0 calculated: N 17.71; found:N 17.67.

EXAMPLE 3 Preparation of the product of formula 1 wherein:

R H; R; c|-i..cH. .c|-i.oH n m 200 Doty's curve).

250 (ultracentrifuge) (average values). The procedure was identical tothat of Example No. 1 but with the substitution of propanolamine forethanolamine.

Yield: about 42 g.; Analysis: for (Cd-1 N 0 calculated: N 16.27; found:N 16.18.

EXAMPLE 4 Preparation of the product of formula I wherein:

R H; R; -CH CH(OH)CH:, n m 200 (Dotys curve EXAMPLE 5 Preparation of theproduct of formula I wherein:

1 CH:; R c HQCH-JOH n m 200 (Doty's curve). 250 (ultracentrifuge)(average values). 5 g of anhydropolyaspartic acid were dissolved in 25ml of 4 dimethylformamide; then 50 ml of Z-methylaminoethanol were addedwithout cooling. A large precipitate formed immediately, whichredissolved during the procedure of raction. After /2 hour, the solutionobtained was treated with acetone and a pasty mass was obtained, thenwashed with acetone, and thereafter treated as in Example No. 3. Yield:about 5 g.; Analysis: for (C H N O calculated: N 16.27; found: N 15.91.

EXAMPLE 6 Preparation of the product of formula 1 wherein:

R, R CH- CH -OH n m 200 (Dotys curve);

Yield: about 7 g.; Analysis: for (Cd-1 N 0 calculated: N 13.85; found: N13.80.

EXAMPLE 7 lnfusional solution for intravenous administration,characterized by the following composition per cent:

a.B-poly-DL-aspartic hydroxyethylamides 3 g Sodium Chloride 0.83 gPotassium Chloride 0.04 g Calcium Chloride 0.065 g Sterile apyrogenicdistilled water as required to ml The solution is prepared in bottles of250-500-1000 ml. Indications: hypovolaemic shock due to haemorrhage ofvarious types, such as trauma, sepsis, poisoning,

burns.

EXAMPLE 8 Infusional solution for intravenous administration,characterized by the following composition per cent:

aJlpolyaspartic hydroxyalkylamides 2.5 4.5 g Sodium chloride 0.83 0.85 gPotassium chloride 0.035 0.04 g Calcium chloride 0.06 0.08 g Sterileapyrogenic distilled water as required to 100 ml.

The solution is prepared in bottles of 250-500-1000 ml. Indications:hypovolaemic shock due to haemorrhage of various types such as trauma,sepsis, poisoning, burns.

What is claimed is:

l. A method for the treatment of a patient with a diminished plasmavolume level and microcirculatory impairment due to septic or traumaticshock, said method comprising intravenously administrating to thepatient a therapeutically effective plasma expanding dose of a compoundof a hydroxyalkylamide of a,B--

polyas partic acid having the formula wherein: R is hydrogen, an alkylgroup, or a hydroxyalkyl group; R is a hydroxyalkyl group; n is thenumber 4. A method as claimed in claim 3 wherein said compound ispresent in an amount of 34% in said solution. 5. A method as claimed inclaim 4 wherein said solution is a saline solution.

1. A METHOD FOR THE TREATMENT OF A PATIENT WITH A DIMINISHED PLASMA VOLUME LEVEL AND MIRCROCIRCULATORY IMPAIRMENT DUE TO SEPTIC OR TRAUMATIC SHOCK, SAID METHOD COMPRISING INTRAVENOUSLY ADMINISTRATING TO THE PATIENT A THERAPEUTICALLY EFFECTIVE PLASMA EXPANDING DOSE OF A COMPOUND OF A HYDROXYALKYLAMIDE OF A,B-POLYASPARTIC ACID HAVING THE FORMULA
 2. A method as claimed in claim 1 wherein said compound is the hydroxyethylamide of Alpha , Beta -poly-DL-aspartic acid.
 3. A method as claimed in claim 1 wherein said compound is in association with a pharmaceutical carrier therefor.
 4. A method as claimed in claim 3 wherein said compound is present in an amount of 3-4% in said solution.
 5. A method as claimed in claim 4 wherein said solution is a saline solution. 